Researchers can query the DNA methylation status of specific sequences within a CpG island using methylation-specific PCR (MSP), a powerful, quick and cost-effective method accessible to virtually any PCR-equipped lab. The principle is simple. First, sodium bisulfite is used to deaminate unmethylat...
We have extended this principle and developed a novel approach, called methylation-specific oligonucleotide (MSO) microarray, for detecting changes of DNA methylation in cancer. The method uses bisulfite-modified DNA as a template for PCR amplification, resulting in conversion of unmethylat...
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Briefly, the first round of PCR was conducted using one bead containing bisulfite-treated DNA with the outer primers and Taq HS premix (TaKaRa). For the second round of PCR, MS-PCR primers and 2 µl of the first PCR amplified products were used. To identify the specific amplification ...
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A systematic comparison of quantitative high-resolution DNA methylation analysis and methylation-specific PCR. Epigenetics. 2012;7(7):772–80. Article CAS PubMed PubMed Central Google Scholar Quillien V, Lavenu A, Karayan-Tapon L, Carpentier C, Labussiere M, Lesimple T, et al. Comparative...
The specific methylation targets were validated by pyrosequencing, Targeted Bisulfite Sequencing Assay, and Multiplex Methylation PCR Assay(MMPA). Results cg26977936, cg23351954, and cg27209741 were validated as differentially methylated and showed the potential diagnostic values (sensitivity and specificity ...
Here, we show that cell type and cancer-specific methylation changes can also be detected, as well as cancer-associated fragmentation signatures. This feasibility study suggests that ONT shallow WGS could be a powerful tool for liquid biopsy. Graphical Abstract...
All the experiments were performed according to protocols described previously [18]. The primer pairs for the methylation specific PCR were either adopted (APC, BRCA1, CDH1, DAPK1, hMLH1, p14ARF, p15INK4b, RASSF1A, RB1andVHL) or designed according to the same principle with assistance of ...
Bisulfite treatment results in a less specific DNA strand for the primers. To compensate for lower specificity, (semi-)nested-PCR was performed in two successive runs with two different sets of primer pairs (one outer primer pair and one inner primer pair). Nested-PCR was followed by an aut...